Fluorescence spectroscopy of single biomolecules

Weiss S 1999 Fluorescence spectroscopy of single biomolecules Science 283 1676-83... 

Weiss S. Fluorescence spectroscopy of single biomolecules. Science 1999, 283, 1676-1683. 

S. Weiss, Fluorescence Spectroscopy of Single Biomolecules , Science, 283,1676 (1999) W.E. Moerner, M. Orrit, Illuminating Single Molecules in Condensed Matter , Science, 283, 1670 (1999)... 

Figure 4.34 Fluorescence spectroscopy of single biomolecules (A) Localization of a macromolecule labeled with a single fluorophore F with nanometer accuracy. The point-spread-function (PSF) can be...

Weiss, S., Fluorescence spectroscopy of single biomolecules. Science, 1999. 283(5408) p. 1676-1683. Xie, X.S., Single-Molecule Spectroscopy and Dynamics At Room Temperature. Accounts of Chemical Research, 1996. 29(12) p. 598-606. 

Weiss, S. 2000. Measuring conformational dynamics of biomolecules by single molecule fluorescence spectroscopy. Nat. Struct. Biol. 7 724-729. 

In recent years, it has become possible to combine fluorescence spectroscopy with optical microscopy to produce localized images of fluorophores in complex matrices such as single cells. In some cases the intrinsic (native) fluorescence of biomolecules can be used in conjunction with microscopy to monitor dynamics in cells. In the absence of a native fluorophore, fluorescent indicators can be used to probe biological events. A particularly interesting probe is the so-called ion probe that changes its excitation or emission spectrum on binding to specific ions such as Ca or Na. These indicators can be used to record events that take place in different parts of individual neurons or to monitor simultaneously the activity of a collection of neurons. 

In recent years, a new approach to condensed-phase spectroscopy has emerged, that focuses on the spectral properties of a single molecule (SM) embedded in a condensed phase [14], Thanks to experimental advances made in optics and microscopy [5], it is now possible to perform single molecule spectroscopy (SMS) in many different systems. Motivations for SMS arise from a fundamental point of view (e.g., the investigation of the field-matter interaction at the level of a SM and the verification of statistical assumptions made in ensemble spectroscopy) and from the possibility of applications (e.g., the use of SMS as a probe for large biomolecules for which a SM is attached as a fluorescent marker). 

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